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Plectin通过诱导F-actin聚合增强肝癌细胞的迁移能力

Plectin Promotes the Migration of Hepatocellular Carcinoma Cells Through Inducing F-actin Polymerization

    摘要
  • 摘要:
    目的 研究Plectin的表达与肝癌细胞迁移能力的关系，揭示Plectin表达影响肝癌细胞迁移行为的分子机理。
    方法 首先，Western blot检测正常肝细胞和肝癌细胞中Plectin的表达。其次，构建Plectin下调的肝癌细胞株，设立对照组（shNC组）和shPLEC组，各组分别设溶剂对照组（shNC+DMSO组或shPLEC+DMSO组）和F-actin骨架聚合诱导剂Jasplakinolide组（shNC+Jasp组或shPLEC+Jasp组）。采用Western blot检测各组肝癌细胞中Plectin的表达及上皮-间质转化（epithelial-mesenchymal transition, EMT）相关分子（N-cadherin、vimentin和E-cadherin）的表达；采用Transwell小室法分析肝癌细胞的迁移能力；采用KEGG（Kyoto Encyclopedia of Genes and Genomes）分析与Plectin基因有关的信号通路；采用免疫荧光技术检测Plectin表达变化对细胞骨架F-actin聚合的影响。
    结果 与正常肝细胞相比，Plectin在肝癌细胞中高表达。与shNC组相比，shPLEC组Plectin的表达降低（P<0.05），肝癌细胞的迁移能力减弱（P<0.05），EMT进程被抑制（N-cadherin和vimentin表达降低，E-cadherin表达升高）（P<0.05）；KEGG分析发现细胞骨架F-actin调控与Plectin的联系最为密切，shPLEC组肝癌细胞骨架F-actin发生解聚。采用F-actin骨架聚合诱导剂Jasplakinolide处理后，与shPLEC+DMSO组相比，shPLEC+Jasp组肝癌细胞迁移能力增强（P<0.05），EMT进程有所恢复（N-cadherin和vimentin表达升高，E-cadherin表达降低）（P<0.05），同时肝癌细胞骨架F-actin聚合亦有所恢复。
    结论 Plectin在肝癌细胞中高表达，肝癌细胞中Plectin通过诱导F-actin聚合促进肝癌细胞的迁移和EMT。

     

    Abstract:
    Objective To explore the relationship between the expression of plectin and the migration of hepatocellular carcinoma (HCC) cells and to elucidate the molecular mechanisms by which plectin expression affects the migration of HCC cells.
    Methods  First of all, Western blot was performed to determine the expression of plectin in normal hepatocytes and HCC cells. Secondly, a plectin-downregulated HCC cell strain was established and the control group (shNC group) and shPLEC group were set up. Each group was divided into a vehicle control group (shNC+DMSO group or shPLEC+DMSO group) and a F-actin cytoskeleton polymerization inducer Jasplakinolide group (shNC+Jasp group or shPLEC+Jasp group). Western blot was performed to determine the expression of plectin and epithelial-mesenchymal transition (EMT)-related proteins, including N-cadherin, vimentin, and E-cadherin. HCC cell migration was evaluated by Transwell assay. KEGG (Kyoto Encyclopedia of Genes and Genomes) was used to analyze the signaling pathways related to plectin gene. The polymerization of F-actin was analyzed by immunofluorescence assay.
    Results  Compared with the normal hepatocytes, HCC cells showed high expression of plectin. Compared with those in the shNC group, the expression of plectin in the shPLEC group was decreased (P<0.05), the migration ability of HCC cells was weakened (P<0.05), and the EMT process was inhibited (with the expression of N-cadherin and vimentin being decreased and the expression of E-cadherin being increased) (P<0.05). KEGG analysis showed that the regulation of cytoskeletal F-actin was most closely associated with plectin and cytoskeletal F-actin depolymerized in the shPLEC group. After treatment with Jasplakinolide, an inducer of F-actin cytoskeleton polymerization, the migration ability of HCC cells in the shPLEC+Jasp group was enhanced compared with that of shPLEC+DMSO group (P<0.05) and the EMT process was restored (with the expression of N-cadherin and vimentin being increased and the expression of E-cadherin being decreased) (P<0.05). In addition, the polymerization of cytoskeletal F-actin in HCC cells was also restored.
    Conclusion  Plectin is highly expressed in HCC cells. Plectin promotes the migration and the EMT of HCC cells through inducing F-actin polymerization.

     

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